Chlamydial infections represent a major threat to the long-term survival of the koala and a successful vaccine would provide a valuable management tool. Vaccination however has the potential to enhance inflammatory disease in animals exposed to a natural infection prior to vaccination, a finding in early human and primate trials of whole cell vaccines to prevent trachoma. In the present study, we vaccinated both healthy koalas as well as clinically diseased koalas with a multi-subunit vaccine consisting of Chlamydia pecorum MOMP and NrdB mixed with immune stimulating complex as adjuvant. Following vaccination, there was no increase in inflammatory pathological changes in animals previously infected with Chlamydia. Strong antibody (including neutralizing antibodies) and lymphocyte proliferation responses were recorded in all vaccinated koalas, both healthy and clinically diseased. Vaccine induced antibodies specific for both vaccine antigens were observed not only in plasma but also in ocular secretions. Our data shows that an experimental chlamydial vaccine is safe to use in previously infected koalas, in that it does not worsen infection-associated lesions. Furthermore, the prototype vaccine is effective, as demonstrated by strong levels of neutralizing antibody and lymphocyte proliferation responses in both healthy and clinically diseased koalas. Collectively, this work illustrates the feasibility of developing a safe and effective Chlamydia vaccine as a tool for management of disease in wild koalas.

Complex interactions between Chlamydia pecorum infection, the immune response and disease exist in the koala. We used quantitative polymerase chain reaction to investigate the relationship between C. pecorum infectious load and ocular and urogenital tract disease. Chlamydia pecorum shedding was generally higher in animals with chronic, active disease than in animals with inactive disease. The absence of ocular disease was generally associated with low levels of shedding, but relatively high levels of shedding in the urogenital tract were detected in some koalas without clinical disease signs. These results suggest a complex disease pathogenesis and clinical course in C. pecorum-infected koalas.

Management actions designed to mitigate development or anthropogenic impacts on species of conservation concern are often implemented without quantifying the benefit to the species. It is often unclear what combinations and intensities of management actions are required to achieve meaningful conservation outcomes. We investigate whether disease and predator control can reverse population declines of koalas (Phascolarctos cinereus). Based on longitudinal monitoring of the epidemiological and demographic status of over 500 animals over 4 years, coupled with an intensive disease and predator management programme, we use survival analyses to estimate annual age‐specific survival rates and population growth, and simulations to quantify the benefit of these actions. Predation and disease accounted for 63% and 29% of mortality, respectively, across all years, with wild dog (dingoes or dingo‐hybrids: Canis dingo, C. dingo × Canis familiaris), carpet pythons (Morelia spilota) and domestic dogs (C. familiaris) accounting for 82%, 14% and 4% of confirmed predation mortalities, respectively. In the first 2 years, before disease and dog control had major impact, the population was declining rapidly with annual growth rates of 0.66 and 0.90. In the third and fourth years, after interventions had been fully implemented, the population growth rate had increased to 1.08 and 1.20. The intrinsic survival rate of joeys was 71.2% (excluding deaths resulting from the death of the mother). Adult survival rates varied as a function of sex, age and year. Even in a declining koala population, management actions can achieve meaningful conservation outcomes (population growth rates greater than one). However, benefits may be short‐lived in the absence of longer term strategies to manage threats. This work also identifies wild dogs as a major threat to koalas, highlighting the need to better understand how wild dog impacts vary in space and time. Policy implications. Offsetting policy that addresses habitat loss alone may achieve little or no meaningful benefit to declining koalas populations. Management must address suites of threats affecting these populations and ensure that the cumulative effects of these actions achieve positive population growth rates.

Chlamydial infections in koalas can cause life-threatening diseases leading to blindness and sterility. However, little is known about the systemic spread of chlamydiae in the inner organs of the koala, and data concerning related pathological organ lesions are limited. The aim of this study was to perform a thorough investigation of organs from 23 koalas and to correlate their histopathological lesions to molecular chlamydial detection. To reach this goal, 246 formalin-fixed and paraffin embedded organ samples from 23 koalas were investigated by histopathology, Chlamydiaceae real-time PCR and immunohistochemistry, ArrayTube Microarray for Chlamydiaceae species identification as well as Chlamydiales real-time PCR and sequencing. By PCR, two koalas were positive for Chlamydia pecorum whereas immunohistochemical labelling for Chlamydiaceae was detected in 10 tissues out of nine koalas. The majority of these (n=6) had positive labelling in the urogenital tract related to histopathological lesions such as cystitis, endometritis, pyelonephritis and prostatitis. Somehow unexpected was the positive labelling in the gastrointestinal tract including the cloaca as well as in lung and spleen indicating systemic spread of infection. Uncultured Chlamydiales were detected in several organs of seven koalas by PCR, and four of these suffered from plasmacytic enteritis of unknown aetiology. Whether the finding of Chlamydia-like organisms in the gastrointestinal tract is linked to plasmacytic enteritis is unclear and remains speculative. However, as recently shown in a mouse model, the gastrointestinal tract might play a role being the site for persistent chlamydial infections and being a source for reinfection of the genital tract. Chlamydial infections in koalas can cause life-threatening diseases leading to blindness and 28 sterility. However, little is known about the systemic spread of chlamydiae in the inner organs of 29 the koala, and data concerning related pathological organ lesions are limited. The aim of this study 30 was to perform a thorough investigation of organs from 23 koalas and to correlate their 31 histopathological lesions to molecular chlamydial detection. To reach this goal, 246 formalin-fixed 32 and paraffin embedded organ samples from 23 koalas were investigated by histopathology, 33Chlamydiaceae real-time PCR and immunohistochemistry, ArrayTube Microarray for 34Chlamydiaceae species identification as well as Chlamydiales real-time PCR and sequencing. By 35 PCR, two koalas were positive for Chlamydia pecorum whereas immunohistochemical labelling for 36Chlamydiaceae was detected in 10 tissues out of nine koalas. The majority of these (n=6)

Simple SummaryWe designed a programme to control free-ranging kangaroos on a Queensland golf course, using contraceptive implants in females and vasectomisation or testicle removal in males. This reduced the numbers of pouch young to about one half of pre-intervention levels and controlled the population over a 2–4 year period. However, the necessary darting caused a mortality rate of 5–10% of captured animals, mainly due to complications before and after anaesthesia. It is concluded that population control is possible but careful management of kangaroos around the time of anaesthesia induction and recovery is important in such programmes to minimise losses.AbstractTraditionally, culling has been the expedient, most common, and in many cases, the only tool used to control free-ranging kangaroo populations. We applied a reproductive control program to a population of eastern grey kangaroos confined to a golf course in South East Queensland. The program aimed to reduce fecundity sufficiently for the population to decrease over time so that overgrazing of the fairways and the frequency of human–animal conflict situations were minimised. In 2003, 92% of the female kangaroos above 5 kg bodyweight were implanted with the GnRH agonist deslorelin after darting with a dissociative anaesthetic. In 2007, 86% of the females above 5 kg were implanted with deslorelin and also 87% of the males above 5 kg were sterilised by either orchidectomy or vasectomy. In 2005, 2008 and 2009, the population was censused to assess the effect of each treatment. The 2003 deslorelin program resulted in effective zero population growth for approximately 2.5 years. The combined deslorelin–surgery program in 2007 reduced the birth rate from 0.3 to 0.06%/year for 16 months, resulting in a 27% population reduction by November 2009. The results were consistent with implants conferring contraception to 100% of implanted females for at least 12 months. The iatrogenic mortality rates for each program were 10.5% and 4.9%, respectively, with 50% of all mortalities due to darting-related injuries, exertional myopathy/hyperthermia or recovery misadventure. The short term sexual and agonistic behaviour of the males was assessed for the 2007 program: no significant changes were seen in adult males given the vasectomy procedure, while sexual behaviours’ were decreased in adult males given the orchidectomy procedure. It is concluded that female reproduction was effectively controlled by implantation with deslorrelin and male reproductive behaviour was reduced by orchidectomy, which together achieved population control.